BRENDA - Enzyme Database show
show all sequences of 2.7.1.49

Overexpression, purification and characterization of two pyrimidine kinases involved in the biosynthesis of thiamin: 4-amino-5-hydroxymethyl-2-methylpyrimidine kinase and 4-amino-5-hydroxymethyl-2-methylpyrimidine phosphate kinase

Reddick, J.J.; Kinsland, C.; Nicewonger, R.; Christian, T.; Downs, D.M.; Winkler, M.E.; Begley, T.P.; Tetrahedron 54, 15983-15991 (1998)
No PubMed abstract available

Data extracted from this reference:

Application
Application
Commentary
Organism
synthesis
useful reagent for the preparation of intermediates on the thiamin biosynthetic pathway
Escherichia coli
Cloned(Commentary)
Commentary
Organism
pET overexpression system
Escherichia coli
KM Value [mM]
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0285
-
pyridoxine
pH 8.0, 25°C
Escherichia coli
0.0335
-
2-methyl-4-amino-5-hydroxymethylpyrimidine
pH 8.0, 25°C
Escherichia coli
0.1431
-
ATP
pH 8.0, 25°C
Escherichia coli
Molecular Weight [Da]
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
gel filtration indicates the enzyme is tetrameric in the native state
Escherichia coli
33000
-
4 * 33000, SDS-PAGE
Escherichia coli
33534
-
4 * 33534, predicted from amino acid sequence
Escherichia coli
Natural Substrates/ Products (Substrates)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine
Escherichia coli
involved in thiamine biosynthetic pathway, first steps
ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
Escherichia coli
?
Organism
Organism
Primary Accession No. (UniProt)
Commentary
Textmining
Escherichia coli
-
BL21(DE3)
-
Purification (Commentary)
Commentary
Organism
-
Escherichia coli
Storage Stability
Storage Stability
Organism
-70°C, stored in 10% glycerol, loses half of its activity after 1 month
Escherichia coli
Substrates and Products (Substrate)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine
-
641660
Escherichia coli
ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
641660
Escherichia coli
?
ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine
involved in thiamine biosynthetic pathway, first steps
641660
Escherichia coli
ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
641660
Escherichia coli
?
ATP + pyridoxine
-
641660
Escherichia coli
ADP + pyridoxine 5'-phosphate
-
641660
Escherichia coli
?
additional information
enzyme is probably different from previously isolated HMP kinase, because the subunit molecular masses are significantly different
641660
Escherichia coli
?
-
-
-
-
Subunits
Subunits
Commentary
Organism
tetramer
4 * 33000, SDS-PAGE; 4 * 33534, predicted from amino acid sequence
Escherichia coli
Turnover Number [1/s]
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.36
-
pyridoxine
pH 8.0, 25°C
Escherichia coli
0.43
-
2-methyl-4-amino-5-hydroxymethylpyrimidine
pH 8.0, 25°C
Escherichia coli
0.43
-
ATP
pH 8.0, 25°C
Escherichia coli
Application (protein specific)
Application
Commentary
Organism
synthesis
useful reagent for the preparation of intermediates on the thiamin biosynthetic pathway
Escherichia coli
Cloned(Commentary) (protein specific)
Commentary
Organism
pET overexpression system
Escherichia coli
KM Value [mM] (protein specific)
KM Value [mM]
KM Value Maximum [mM]
Substrate
Commentary
Organism
Structure
0.0285
-
pyridoxine
pH 8.0, 25°C
Escherichia coli
0.0335
-
2-methyl-4-amino-5-hydroxymethylpyrimidine
pH 8.0, 25°C
Escherichia coli
0.1431
-
ATP
pH 8.0, 25°C
Escherichia coli
Molecular Weight [Da] (protein specific)
Molecular Weight [Da]
Molecular Weight Maximum [Da]
Commentary
Organism
additional information
-
gel filtration indicates the enzyme is tetrameric in the native state
Escherichia coli
33000
-
4 * 33000, SDS-PAGE
Escherichia coli
33534
-
4 * 33534, predicted from amino acid sequence
Escherichia coli
Natural Substrates/ Products (Substrates) (protein specific)
Natural Substrates
Organism
Commentary (Nat. Sub.)
Natural Products
Commentary (Nat. Pro.)
Organism (Nat. Pro.)
Reversibility
ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine
Escherichia coli
involved in thiamine biosynthetic pathway, first steps
ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
Escherichia coli
?
Purification (Commentary) (protein specific)
Commentary
Organism
-
Escherichia coli
Storage Stability (protein specific)
Storage Stability
Organism
-70°C, stored in 10% glycerol, loses half of its activity after 1 month
Escherichia coli
Substrates and Products (Substrate) (protein specific)
Substrates
Commentary Substrates
Literature (Substrates)
Organism
Products
Commentary (Products)
Literature (Products)
Organism (Products)
Reversibility
ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine
-
641660
Escherichia coli
ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
641660
Escherichia coli
?
ATP + 2-methyl-4-amino-5-hydroxymethylpyrimidine
involved in thiamine biosynthetic pathway, first steps
641660
Escherichia coli
ADP + 2-methyl-4-amino-5-phosphomethylpyrimidine
-
641660
Escherichia coli
?
ATP + pyridoxine
-
641660
Escherichia coli
ADP + pyridoxine 5'-phosphate
-
641660
Escherichia coli
?
additional information
enzyme is probably different from previously isolated HMP kinase, because the subunit molecular masses are significantly different
641660
Escherichia coli
?
-
-
-
-
Subunits (protein specific)
Subunits
Commentary
Organism
tetramer
4 * 33000, SDS-PAGE; 4 * 33534, predicted from amino acid sequence
Escherichia coli
Turnover Number [1/s] (protein specific)
Turnover Number Minimum [1/s]
Turnover Number Maximum [1/s]
Substrate
Commentary
Organism
Structure
0.36
-
pyridoxine
pH 8.0, 25°C
Escherichia coli
0.43
-
2-methyl-4-amino-5-hydroxymethylpyrimidine
pH 8.0, 25°C
Escherichia coli
0.43
-
ATP
pH 8.0, 25°C
Escherichia coli
Other publictions for EC 2.7.1.49
No.
1st author
Pub Med
title
organims
journal
volume
pages
year
Activating Compound
Application
Cloned(Commentary)
Crystallization (Commentary)
Engineering
General Stability
Inhibitors
KM Value [mM]
Localization
Metals/Ions
Molecular Weight [Da]
Natural Substrates/ Products (Substrates)
Organic Solvent Stability
Organism
Oxidation Stability
Posttranslational Modification
Purification (Commentary)
Reaction
Renatured (Commentary)
Source Tissue
Specific Activity [micromol/min/mg]
Storage Stability
Substrates and Products (Substrate)
Subunits
Temperature Optimum [°C]
Temperature Range [°C]
Temperature Stability [°C]
Turnover Number [1/s]
pH Optimum
pH Range
pH Stability
Cofactor
Ki Value [mM]
pI Value
IC50 Value
Activating Compound (protein specific)
Application (protein specific)
Cloned(Commentary) (protein specific)
Cofactor (protein specific)
Crystallization (Commentary) (protein specific)
Engineering (protein specific)
General Stability (protein specific)
IC50 Value (protein specific)
Inhibitors (protein specific)
Ki Value [mM] (protein specific)
KM Value [mM] (protein specific)
Localization (protein specific)
Metals/Ions (protein specific)
Molecular Weight [Da] (protein specific)
Natural Substrates/ Products (Substrates) (protein specific)
Organic Solvent Stability (protein specific)
Oxidation Stability (protein specific)
Posttranslational Modification (protein specific)
Purification (Commentary) (protein specific)
Renatured (Commentary) (protein specific)
Source Tissue (protein specific)
Specific Activity [micromol/min/mg] (protein specific)
Storage Stability (protein specific)
Substrates and Products (Substrate) (protein specific)
Subunits (protein specific)
Temperature Optimum [°C] (protein specific)
Temperature Range [°C] (protein specific)
Temperature Stability [°C] (protein specific)
Turnover Number [1/s] (protein specific)
pH Optimum (protein specific)
pH Range (protein specific)
pH Stability (protein specific)
pI Value (protein specific)
Expression
General Information
General Information (protein specific)
Expression (protein specific)
KCat/KM [mM/s]
KCat/KM [mM/s] (protein specific)
738234
Castro-Fernandez
Emergence of pyridoxal phospho ...
Salmonella enterica subsp. enterica serovar Typhimurium, Salmonella enterica subsp. enterica serovar Typhimurium ATCC 700720
FEBS Lett.
588
3068-3073
2014
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1
1
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738451
Nodwell
A subfamily of bacterial ribok ...
Staphylococcus aureus, Staphylococcus aureus ATCC 700699
J. Am. Chem. Soc.
136
4992-4999
2014
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1
1
2
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1
6
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2
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4
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6
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2
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6
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4
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6
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722048
Nodwell
Rugulactone and its analogues ...
Staphylococcus aureus
ChemBioChem
13
1439-1446
2012
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3
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1
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3
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718477
French
Structure of trifunctional THI ...
Saccharomyces cerevisiae
Acta Crystallogr. Sect. D
67
784-791
2011
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1
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2
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686824
Onozuka
Involvement of thiaminase II e ...
Saccharomyces cerevisiae
FEMS Yeast Res.
8
266-275
2008
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-
1
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1
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1
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1
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2
-
1
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1
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1
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1
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1
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2
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1
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1
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671903
Rapala-Kozik
Molecular characterization of ...
Zea mays
Biochem. J.
408
149-159
2007
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1
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7
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3
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5
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1
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1
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1
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1
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7
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3
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1
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1
1
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1
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672477
Wrenger
Vitamin B1 de novo synthesis i ...
Plasmodium falciparum
Biol. Chem.
387
41-51
2006
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1
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2
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1
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1
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1
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2
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1
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1
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2
-
4
1
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-
1
-
-
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-
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-
-
-
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674302
Karunakaran
Thiamine is synthesized by a s ...
Rhizobium leguminosarum
J. Bacteriol.
188
6661-6668
2006
-
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1
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1
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661316
Haas
Thi20, a remarkable enzyme fro ...
Saccharomyces cerevisiae
Bioorg. Chem.
33
338-344
2005
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1
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1
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3
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1
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1
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1
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1
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1
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661537
Kawasaki
Biosynthesis of hydroxymethylp ...
Saccharomyces cerevisiae, Saccharomyces cerevisiae YPH500
Curr. Genet.
47
156-162
2005
-
-
1
-
1
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1
1
4
-
4
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1
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1
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6
1
1
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1
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1
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1
1
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1
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1
1
4
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1
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1
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6
1
1
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1
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-
662007
Park
Characterization of two kinase ...
Bacillus subtilis
J. Bacteriol.
186
1571-1573
2004
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1
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1
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1
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3
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2
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1
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4
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1
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1
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3
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1
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4
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1
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1
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641661
Mizote
Cloning and characterization o ...
Escherichia coli, Escherichia coli BL21(DE3) (pLysS)
Microbiology
145
495-501
1999
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1
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2
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1
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1
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639726
Kim
A Brassica cDNA clone encoding ...
Arabidopsis thaliana, Brassica napus
Plant Mol. Biol.
37
955-966
1998
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2
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641660
Reddick
-
Overexpression, purification a ...
Escherichia coli
Tetrahedron
54
15983-15991
1998
-
1
1
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-
3
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3
1
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1
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1
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4
1
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1
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1
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641658
Mizote
Purification and properties of ...
Escherichia coli
Biochim. Biophys. Acta
991
109-113
1989
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2
4
1
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1
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5
1
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2
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5
1
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2
2
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641657
Lewin
-
The biosynthesis of thiamine. ...
Saccharomyces cerevisiae
J. Biol. Chem.
236
2768-2771
1961
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